CN108017720A - Flower of JINHUAKUI extraction method of polysaccharides - Google Patents
Flower of JINHUAKUI extraction method of polysaccharides Download PDFInfo
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- CN108017720A CN108017720A CN201810114050.0A CN201810114050A CN108017720A CN 108017720 A CN108017720 A CN 108017720A CN 201810114050 A CN201810114050 A CN 201810114050A CN 108017720 A CN108017720 A CN 108017720A
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- jinhuakui
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- 238000000605 extraction Methods 0.000 title claims abstract description 106
- 150000004676 glycans Chemical class 0.000 title claims abstract description 105
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 105
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 105
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 97
- 239000000706 filtrate Substances 0.000 claims abstract description 59
- 235000019441 ethanol Nutrition 0.000 claims abstract description 45
- 239000000284 extract Substances 0.000 claims abstract description 42
- 239000007788 liquid Substances 0.000 claims abstract description 42
- 238000001914 filtration Methods 0.000 claims abstract description 31
- 238000001035 drying Methods 0.000 claims abstract description 27
- 239000013049 sediment Substances 0.000 claims abstract description 26
- 238000001556 precipitation Methods 0.000 claims abstract description 24
- 238000003756 stirring Methods 0.000 claims abstract description 24
- 238000007789 sealing Methods 0.000 claims abstract description 16
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims abstract description 15
- 238000010926 purge Methods 0.000 claims abstract description 9
- 108010059892 Cellulase Proteins 0.000 claims description 42
- 229940106157 cellulase Drugs 0.000 claims description 42
- 238000006243 chemical reaction Methods 0.000 claims description 38
- 239000012153 distilled water Substances 0.000 claims description 34
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 34
- 238000010438 heat treatment Methods 0.000 claims description 16
- 230000007246 mechanism Effects 0.000 claims description 15
- 239000012530 fluid Substances 0.000 claims description 13
- 238000005119 centrifugation Methods 0.000 claims description 10
- 210000004700 fetal blood Anatomy 0.000 claims description 9
- 108010084185 Cellulases Proteins 0.000 claims description 7
- 102000005575 Cellulases Human genes 0.000 claims description 7
- 238000004891 communication Methods 0.000 claims description 7
- 239000011229 interlayer Substances 0.000 claims description 6
- 230000002045 lasting effect Effects 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 3
- 239000002244 precipitate Substances 0.000 claims description 3
- 206010013786 Dry skin Diseases 0.000 claims 1
- 230000004044 response Effects 0.000 abstract description 20
- 238000000034 method Methods 0.000 abstract description 18
- 238000011084 recovery Methods 0.000 abstract description 10
- 230000011218 segmentation Effects 0.000 abstract 1
- 108090000790 Enzymes Proteins 0.000 description 26
- 102000004190 Enzymes Human genes 0.000 description 26
- 229940088598 enzyme Drugs 0.000 description 26
- 238000002474 experimental method Methods 0.000 description 19
- 241000196324 Embryophyta Species 0.000 description 7
- 210000004027 cell Anatomy 0.000 description 7
- 238000013401 experimental design Methods 0.000 description 7
- 238000013461 design Methods 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 238000005457 optimization Methods 0.000 description 4
- 230000008569 process Effects 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 244000020551 Helianthus annuus Species 0.000 description 3
- 235000003222 Helianthus annuus Nutrition 0.000 description 3
- 238000000540 analysis of variance Methods 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 229920002678 cellulose Polymers 0.000 description 3
- 239000001913 cellulose Substances 0.000 description 3
- 238000004140 cleaning Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 230000002779 inactivation Effects 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 235000011351 tree mallow Nutrition 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- FFRBMBIXVSCUFS-UHFFFAOYSA-N 2,4-dinitro-1-naphthol Chemical compound C1=CC=C2C(O)=C([N+]([O-])=O)C=C([N+]([O-])=O)C2=C1 FFRBMBIXVSCUFS-UHFFFAOYSA-N 0.000 description 1
- 241001075517 Abelmoschus Species 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 241000881780 Lavatera Species 0.000 description 1
- 240000002853 Nelumbo nucifera Species 0.000 description 1
- 235000006508 Nelumbo nucifera Nutrition 0.000 description 1
- 235000006510 Nelumbo pentapetala Nutrition 0.000 description 1
- 241001409201 Tabebuia chrysantha Species 0.000 description 1
- GAMYVSCDDLXAQW-AOIWZFSPSA-N Thermopsosid Natural products O(C)c1c(O)ccc(C=2Oc3c(c(O)cc(O[C@H]4[C@H](O)[C@@H](O)[C@H](O)[C@H](CO)O4)c3)C(=O)C=2)c1 GAMYVSCDDLXAQW-AOIWZFSPSA-N 0.000 description 1
- 241000707822 Ulmus glabra Species 0.000 description 1
- 229930003427 Vitamin E Natural products 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 229930003944 flavone Natural products 0.000 description 1
- 150000002212 flavone derivatives Chemical class 0.000 description 1
- 235000011949 flavones Nutrition 0.000 description 1
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 230000017525 heat dissipation Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000011344 liquid material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 239000000419 plant extract Substances 0.000 description 1
- 235000019353 potassium silicate Nutrition 0.000 description 1
- 238000000611 regression analysis Methods 0.000 description 1
- NTHWMYGWWRZVTN-UHFFFAOYSA-N sodium silicate Chemical compound [Na+].[Na+].[O-][Si]([O-])=O NTHWMYGWWRZVTN-UHFFFAOYSA-N 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 238000005211 surface analysis Methods 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
- VHBFFQKBGNRLFZ-UHFFFAOYSA-N vitamin p Natural products O1C2=CC=CC=C2C(=O)C=C1C1=CC=CC=C1 VHBFFQKBGNRLFZ-UHFFFAOYSA-N 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Sustainable Development (AREA)
- Biochemistry (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Organic Chemistry (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
The invention discloses a kind of flower of JINHUAKUI extraction method of polysaccharides, comprise the following steps：Drying to constant weight for flower of JINHUAKUI gradient, obtains dry flower of JINHUAKUI, and crushing obtains Golden flower pollen；The enzymolysis liquid that segmentation in obtained Golden flower pollen is added to quantitative constant temperature stirs evenly, and adjusts pH value to after 6.5 7, persistently stirs and extracts, and obtains enzymolysis mixed liquor；Enzymolysis mixed liquor is inactivated and centrifuged, filtrate is placed in extraction intracavitary after filtering；Ethanol is added into filtrate, sealing and standing, obtains alcohol precipitation mixed liquor；Alcohol precipitation mixed liquor is centrifuged, sediment is stayed in filtering；Sediment sequential purge and is precipitated in ethanol that mass concentration is 95%, absolute ethyl alcohol and acetone, then constant temperature drying to constant weight, up to the flower of JINHUAKUI polysaccharide.Its combining response face method Optimized Extraction Process, realizes the rapid extraction of flower of JINHUAKUI polysaccharide, and recovery rate is high, and extraction cost is low.
Description
Technical field
The present invention relates to technical field of polysaccharide extraction, more particularly to a kind of flower of JINHUAKUI extraction method of polysaccharides.
Background technology
In recent years, various circles of society all extremely pay close attention to plant extract research, and numerous plant components are largely extracted simultaneously
Utilize, so that emerging many industries, have developed numerous industrial chains.As the improvement of people's living standards, since polysaccharide has
Very high utility value, the extraction of plant polyose is also into research hotspot.Herbaceous plant Golden flower
(HibiseumanihotL.) Malvaceae Abelmoschus is belonged to, its life cycle was completed in 1 year, according to its growth habit and shape
State feature people are also known as its vegetable Furong, glutinous dry, wych-elm skin and wild lotus etc., and Golden flower is rich in multiclass trace element, natural flavone
The components such as class compound, vitamin E, unrighted acid and the unexistent collagen of other plant, there is very high medicine to eat
Value.The primary efficacy of Golden flower is to reduce feeling of pain, heat dissipation, resist body inflammation, adjust blood fat, improve immunity, prevent
Oxidation, suppress tumour cell etc..Flower of JINHUAKUI, also with ornamental value, both can independently plant, colony in golden yellow in small institute
The row such as plantation, roadside, government department, home for destitute is planted, piece is planted, can also be in the region such as field, park, garden large-scale planting.
In summary, it may be said that Golden flower whole body is all precious, there is wide development prospect.But polysaccharide contained by flower of JINHUAKUI is ground at present
Study carefully rarely found, need to be developed and utilized, and as a kind of endangered plant Golden flower, its contained polysaccharide of current each bound pair
Research is also very rarely seen.
Golden flower is many rich in nutritional ingredient, and nutritive value is high, has very high Development volue.The medicine price cheating of plant polyose
Value, health value etc. are very high, are accepted extensively.Exploitation Golden flower polysaccharide has extensive prospect, and inherently development is made in the future
Medicine, chemical industry, health care, flowers etc. have industry, and pull the rise of new industry.But more sugar types are various, structure is multiple
Miscellaneous, the research for Golden flower polysaccharide is thorough not enough at present, it is still necessary to which researcher is further furtherd investigate.
The content of the invention
It is an object of the invention to solve at least the above, and provide the advantages of at least will be described later.
It is a still further object of the present invention to provide a kind of flower of JINHUAKUI extraction method of polysaccharides, the optimization extraction of combining response face method
Technique, realizes the rapid extraction of flower of JINHUAKUI polysaccharide, and recovery rate is high, and extraction cost is low.
In order to realize these purposes and further advantage according to the present invention, there is provided a kind of flower of JINHUAKUI Polyose extraction side
Method, mainly includes the following steps：
Step 1, flower of JINHUAKUI clean after after 5-10 DEG C of Cord blood 3-5h, be put into be preheated at 55-65 DEG C and extracting
Drying 1-2h in the reaction chamber of device, then reduces reaction chamber temperature and continues to dry 2-4h to 45-55 DEG C, finally reduce reaction chamber temperature
Degree after obtaining dry flower of JINHUAKUI, is crushed to 50-70 mesh, obtains Golden flower pollen to 30-45 DEG C of constant temperature drying to constant weight；
Step 2, by obtained Golden flower pollen according to solid-liquid ratio 1:50-60g/mL weighs quantitative distilled water, Yi Jizhan
The cellulase of the Golden flower pollen gross mass 0.8-1.2%, 3 parts are divided into by the distilled water and cellulase respectively,
And 3 parts of distilled water are separately heated to temperature as 30-45 DEG C, 45-50 DEG C and 60-65 DEG C, and 3 parts of cellulases are distinguished
Add in 3 parts of distilled water to dissolving, obtain 3 portions of cellulase mixed liquors；3 portions of cellulase mixed liquors are then pressed into interval time
20-30min, is added sequentially into the Golden flower pollen and stirs evenly, and adjusts pH value all the time between 6.5-7.5, continues
Stirring, obtains enzymolysis mixed liquor；
Step 3, after the enzymolysis mixed liquor is inactivated 8-12min at a temperature of 85-100 DEG C, in 3800-4200r/
8-12min is centrifuged under the conditions of min, opens the filtrate port of the reaction chamber, the filtrate of the enzymolysis mixed liquor after the centrifugation flows into
The extraction intracavitary of the extractor；
Step 4, the volume according to filtrate in the extractor, the mass concentration of 3-5 times of volume is added into the filtrate
For the ethanol of 92-96%, sealing and standing 10-15h, obtains alcohol precipitation mixed liquor；
Step 5, by the alcohol precipitation mixed liquor centrifuge 10-20min under the conditions of 3800-4200r/min, opens the extraction
Sediment is stayed in the filtering mouth of device, filtering；
Step 6, by the sediment in mass concentration is the ethanol of 92-96%, absolute ethyl alcohol and acetone sequential purge
And precipitate, then constant temperature drying is to constant weight at 55-65 DEG C, up to the flower of JINHUAKUI polysaccharide.
Preferably, in the flower of JINHUAKUI extraction method of polysaccharides, mainly include the following steps：
Step 1, flower of JINHUAKUI clean after after 6 DEG C of Cord blood 4h, be put into and be preheated at 60 DEG C in the reaction of extractor
Intracavitary dries 1.5h, then reduces reaction chamber temperature and continues to dry 3h to 50 DEG C, finally reduces reaction chamber temperature to 35 DEG C of constant temperature
Drying to constant weight, after obtaining dry flower of JINHUAKUI, is crushed to 60 mesh, obtains Golden flower pollen；
Step 2, by obtained Golden flower pollen according to solid-liquid ratio 1:58.28g/mL weighs quantitative distilled water, Yi Jizhan
The cellulase of the Golden flower pollen gross mass 0.95%, 3 parts are divided into by the distilled water and cellulase respectively, and will
3 parts of distilled water are separately heated to temperature as 60 DEG C, 50 DEG C and 65 DEG C, and 3 parts of cellulases are separately added into 3 parts of distilled water
In to dissolve, obtain 3 portions of cellulase mixed liquors；3 portions of cellulase mixed liquors are then pressed into interval time 22min, are sequentially added
Enter into the Golden flower pollen and stir evenly, and it is 7 to adjust pH value all the time, lasting stirring, obtains enzymolysis mixed liquor；
Step 3, after the enzymolysis mixed liquor is inactivated 10min at a temperature of 92 DEG C, centrifuge under the conditions of 4000r/min
10min, opens the filtrate port of the reaction chamber, and the filtrate of the enzymolysis mixed liquor after the centrifugation flows into extraction intracavitary；
Step 4, the volume according to filtrate in the extractor, the mass concentration that 4 times of volumes are added into the filtrate are
95% ethanol, sealing and standing 12h, obtains alcohol precipitation mixed liquor；
Step 5, by the alcohol precipitation mixed liquor centrifuge 15min under the conditions of 4000r/min, opens the filtering of the extractor
Mouthful, sediment is stayed in filtering；
Step 6, sequential purge and sink the sediment in ethanol that mass concentration is 95%, absolute ethyl alcohol and acetone
Form sediment, then constant temperature drying is to constant weight at 60 DEG C, up to the flower of JINHUAKUI polysaccharide.
Preferably, in the flower of JINHUAKUI extraction method of polysaccharides, 60 will be crossed after dry flower of JINHUAKUI crushing in step 1
Mesh sieve.
Preferably, in the flower of JINHUAKUI extraction method of polysaccharides, the mesh of the filter screen of filtrate port is in step 3
65-80 mesh.
Preferably, in the flower of JINHUAKUI extraction method of polysaccharides, the filter screen of the filtering mouth of extractor in step 5
Mesh is 70-95 mesh.
A kind of extractor applied to flower of JINHUAKUI Polyose extraction, including：
Reaction chamber, its upper part opening, the reaction chamber include outer barrel and in the outer barreies and can be with respect to institute
State the rotating inner cylinder of outer barrel；Bottom in the inner cylinder is provided with the first rabbling mechanism, the first rabbling mechanism bag
The first agitating shaft and the first agitating vane are included, the central shaft of first agitating shaft along the inner cylinder is longitudinal through inner cylinder
Inside, first agitating vane are rotated with the first stirring axis connection and by the drive of the first agitating shaft；The outer barrel and
The first motor is additionally provided with interlayer between inner cylinder, is weighed and the first heating film；In the first motor driving is described
Cylinder rotates, and drives first agitating shaft to rotate；Described weigh is arranged at the lower section of the inner cylinder, with to inner cylinder
Interior material is weighed；First heating film is covered on the outer wall of the inner cylinder, to add to the inner cylinder
Heat；Filtrate port is provided with the lower wall of the inner cylinder, the first filter screen is removably provided with the filtrate port；
Extract chamber, its upper part opening, it is described extraction chamber opening on be provided with lid, by it is described extraction chamber opening cover
Lid；The extraction chamber passes through pipeline and the filtrate port fluid communication；The extraction chamber includes outer chamber and installed in described outer
And can the relatively described rotating inner chamber body of outer chamber in cavity；Bottom in the inner chamber body is provided with the second rabbling mechanism, institute
Stating the second rabbling mechanism includes the second agitating shaft and the second agitating vane, central shaft of second agitating shaft along the inner chamber body
Through inside inner chamber body, second agitating vane is revolved with the second stirring axis connection and by the drive of the second agitating shaft for longitudinal direction
Turn；The second motor and the second heating film are additionally provided with interlayer between the outer chamber and inner chamber body；Second motor drives
The dynamic inner chamber body rotation, and drive second agitating shaft to rotate；Second heating film is covered in the outer of the inner chamber body
On wall, to be heated to the inner chamber body；It is provided with filtering mouth in the lower wall of the inner chamber body, it is removable on the filtering mouth
That unloads is provided with the second filter screen.
Preferably, in the extractor, scale is respectively arranged with the inner wall of the inner cylinder and inner chamber body.
Preferably, in the extractor, sealing ring is provided with the lower surface outer rim of the lid, so that the lid
When body is covered in the opening of the extraction chamber, the extraction intracavitary forms sealing space.
Preferably, in the extractor, the intake chute of annular, the drain are additionally provided with below the inner cylinder
Groove downward concave arc centered on setting, so that the intake chute forms the accommodating space of upper opening, the intake chute position
Set in the lower section of the filtrate port, and around the inner cylinder, fluid guidance port, the drain are offered below the intake chute
Mouth and the extraction chamber fluid communication.
The present invention includes at least following beneficial effect：
The present invention by enzyme amount, solid-liquid ratio, three factors of enzymolysis time experiment of single factor, draw flower of JINHUAKUI polysaccharide
Optimum extraction condition.Again with the extraction work of response phase method design Three factors-levels experimental program optimization flower of JINHUAKUI polysaccharide
Skill；Cord blood is placed on by the way that flower of JINHUAKUI is cleaned so that the cell of flower of JINHUAKUI more compacts, then by directly putting
Enter medium temperature environment to be dried so that the loss of intracellular matter substantially reduces, then by declining in gradient at a temperature of into
Row drying, further reduces the loss of flower of JINHUAKUI internal substance, while avoids destruction of the high temperature to internal substance, subsequently
The polysaccharide quality higher of extraction；By the way that quantitative distilled water is divided into 3 parts, and be separately heated to different temperatures and cellulase into
Row mixing so that the enzymolysis liquid of initial lower temperature and dry flower of JINHUAKUI temperature are basically identical, reduce temperature jump to thin
The stimulation of cell wall so that flower of JINHUAKUI digests under gentle adjusting, then adds the slightly higher enzymolysis liquid of temperature, can promote thin
Born of the same parents' broken wall, and polysaccharide is unwind, easy to the extraction of polysaccharide, finally with the enzymolysis liquid of higher temperature, residue can be promoted not extract
Flower of JINHUAKUI breaking-wall cell, enhance contact of the cellulase with cell, effectively raise the recovery rate of polysaccharide so that gold
The extraction rate reached of tree mallow polysaccharide to 4.02%, and by by flower of JINHUAKUI it is dry, crush, enzymolysis, then inactivation, alcohol precipitation,
Filtering, up to flower of JINHUAKUI polysaccharide after cleaning, drying, Polyose extraction step is simple, easy to operate, and cost is low, and Polyose extraction
Rate is high.
Further advantage, target and the feature of the present invention embodies part by following explanation, and part will also be by this
The research and practice of invention and be understood by the person skilled in the art.
Brief description of the drawings
Fig. 1 is the influence figure that enzyme amount extracts flower of JINHUAKUI polysaccharide yield；
Fig. 2 is the influence figure that enzymolysis time extracts flower of JINHUAKUI polysaccharide yield；
Fig. 3 is the influence figure that solid-liquid ratio extracts flower of JINHUAKUI polysaccharide yield；
Fig. 4 is the structure chart of extractor.
Embodiment
The present invention is described in further detail below in conjunction with the accompanying drawings, to make those skilled in the art with reference to specification text
Word can be implemented according to this.
It should be appreciated that such as " having ", "comprising" and " comprising " term used herein are not precluded from one or more
The presence or addition of a other elements or its combination.
The present invention provides a kind of flower of JINHUAKUI extraction method of polysaccharides, mainly includes the following steps：
Step 1, flower of JINHUAKUI clean after after 5-10 DEG C of Cord blood 3-5h, be put into be preheated at 55-65 DEG C and extracting
Drying 1-2h in the reaction chamber of device, then reduces reaction chamber temperature and continues to dry 2-4h to 45-55 DEG C, finally reduce reaction chamber temperature
Degree after obtaining dry flower of JINHUAKUI, is crushed to 50-70 mesh, obtains Golden flower pollen to 30-45 DEG C of constant temperature drying to constant weight；
Step 2, by obtained Golden flower pollen according to solid-liquid ratio 1:50-60g/mL weighs quantitative distilled water, Yi Jizhan
The cellulase of the Golden flower pollen gross mass 0.8-1.2%, 3 parts are divided into by the distilled water and cellulase respectively,
And 3 parts of distilled water are separately heated to temperature as 30-45 DEG C, 45-50 DEG C and 60-65 DEG C, and 3 parts of cellulases are distinguished
Add in 3 parts of distilled water to dissolving, obtain 3 portions of cellulase mixed liquors；3 portions of cellulase mixed liquors are then pressed into interval time
20-30min, is added sequentially into the Golden flower pollen and stirs evenly, and adjusts pH value all the time between 6.5-7.5, continues
Stirring, obtains enzymolysis mixed liquor；
Step 3, after the enzymolysis mixed liquor is inactivated 8-12min at a temperature of 85-100 DEG C, in 3800-4200r/
8-12min is centrifuged under the conditions of min, opens the filtrate port of the reaction chamber, the filtrate of the enzymolysis mixed liquor after the centrifugation flows into
The extraction intracavitary of the extractor；
Step 4, the volume according to filtrate in the extractor, the mass concentration of 3-5 times of volume is added into the filtrate
For the ethanol of 92-96%, sealing and standing 10-15h, obtains alcohol precipitation mixed liquor；
Step 5, by the alcohol precipitation mixed liquor centrifuge 10-20min under the conditions of 3800-4200r/min, opens the extraction
Sediment is stayed in the filtering mouth of device, filtering；
Step 6, by the sediment in mass concentration is the ethanol of 92-96%, absolute ethyl alcohol and acetone sequential purge
And precipitate, then constant temperature drying is to constant weight at 55-65 DEG C, up to the flower of JINHUAKUI polysaccharide.
In such scheme, by enzyme amount, solid-liquid ratio, three factors of enzymolysis time experiment of single factor, draw Golden flower
The optimum extraction condition of flower polysaccharide.Again with response phase method design Three factors-levels experimental program optimization flower of JINHUAKUI polysaccharide
Extraction process；Cord blood is placed on by the way that flower of JINHUAKUI is cleaned so that the cell of flower of JINHUAKUI more compacts, and then passes through
It is directly placed into medium temperature environment to be dried so that the loss of intracellular matter substantially reduces, then the temperature by gradient declining
It is dried under degree, further reduces the loss of flower of JINHUAKUI internal substance, while is avoided high temperature and internal substance is broken
It is bad, the polysaccharide quality higher of subsequent extracted；By the way that quantitative distilled water is divided into 3 parts, and it is separately heated to different temperatures and fibre
The plain enzyme of dimension is mixed so that the enzymolysis liquid of initial lower temperature and dry flower of JINHUAKUI temperature are basically identical, reduce temperature
It is mutated the stimulation to cell membrane so that flower of JINHUAKUI digests under gentle adjusting, then adds the slightly higher enzymolysis liquid of temperature, energy
Enough promote breaking-wall cell, and polysaccharide is unwind, easy to the extraction of polysaccharide, finally with the enzymolysis liquid of higher temperature, can promote to remain
Remaining undrawn flower of JINHUAKUI breaking-wall cell, enhances contact of the cellulase with cell, effectively raises the extraction of polysaccharide
Rate so that the extraction rate reached of flower of JINHUAKUI polysaccharide to 4.02%, and by by flower of JINHUAKUI it is dry, crush, enzymolysis, then go out
Work, alcohol precipitation, filtering, up to flower of JINHUAKUI polysaccharide after cleaning, drying, Polyose extraction step is simple, easy to operate, and cost is low, and
Polysaccharide extract rate is high.
Preferably, in the flower of JINHUAKUI extraction method of polysaccharides, mainly include the following steps：
Step 1, flower of JINHUAKUI clean after after 6 DEG C of Cord blood 4h, be put into and be preheated at 60 DEG C in the reaction of extractor
Intracavitary dries 1.5h, then reduces reaction chamber temperature and continues to dry 3h to 50 DEG C, finally reduces reaction chamber temperature to 35 DEG C of constant temperature
Drying to constant weight, after obtaining dry flower of JINHUAKUI, is crushed to 60 mesh, obtains Golden flower pollen；
Step 2, by obtained Golden flower pollen according to solid-liquid ratio 1:58.28g/mL weighs quantitative distilled water, Yi Jizhan
The cellulase of the Golden flower pollen gross mass 0.95%, 3 parts are divided into by the distilled water and cellulase respectively, and will
3 parts of distilled water are separately heated to temperature as 60 DEG C, 50 DEG C and 65 DEG C, and 3 parts of cellulases are separately added into 3 parts of distilled water
In to dissolve, obtain 3 portions of cellulase mixed liquors；3 portions of cellulase mixed liquors are then pressed into interval time 22min, are sequentially added
Enter into the Golden flower pollen and stir evenly, and it is 7 to adjust pH value all the time, lasting stirring, obtains enzymolysis mixed liquor；
Step 3, after the enzymolysis mixed liquor is inactivated 10min at a temperature of 92 DEG C, centrifuge under the conditions of 4000r/min
10min, opens the filtrate port of the reaction chamber, and the filtrate of the enzymolysis mixed liquor after the centrifugation flows into extraction intracavitary；
Step 4, the volume according to filtrate in the extractor, the mass concentration that 4 times of volumes are added into the filtrate are
95% ethanol, sealing and standing 12h, obtains alcohol precipitation mixed liquor；
Step 5, by the alcohol precipitation mixed liquor centrifuge 15min under the conditions of 4000r/min, opens the filtering of the extractor
Mouthful, sediment is stayed in filtering；
Step 6, sequential purge and sink the sediment in ethanol that mass concentration is 95%, absolute ethyl alcohol and acetone
Form sediment, then constant temperature drying is to constant weight at 60 DEG C, up to the flower of JINHUAKUI polysaccharide.
In one preferred solution, 60 mesh sieves will be crossed after dry flower of JINHUAKUI crushing in step 1.
In such scheme, by the way that the flower of JINHUAKUI after crushing is crossed 60 mesh sieves, it can ensure the flower of JINHUAKUI after crushing
Particle is uniform, easy to be subsequently sufficiently mixed with cellulase, and then improves polysaccharide extract rate.
In one preferred solution, the mesh of the filter screen of filtrate port is 65-80 mesh in step 3.
In such scheme, it is 65-80 mesh by setting the mesh of filter screen of filtrate port, can ensures to be crushed to 60 mesh
Flower of JINHUAKUI will not be by filter screen, and then ensure that obtained filtrate is pure.
In one preferred solution, the mesh of the filter screen of the filtering mouth of extractor is 70-95 mesh in step 5.
In such scheme, it is 70-95 mesh by setting the mesh of filter screen of filtering mouth, can ensures that sediment retains
It is complete, and then reduce flower of JINHUAKUI polysaccharide loss, improve polysaccharide extract rate.
As shown in figure 4, a kind of extractor applied to flower of JINHUAKUI Polyose extraction, including：Reaction chamber 1, its upper part opening,
The reaction chamber 1 is interior including outer barrel 2 and installed in the outer barrel 2 and can the relatively described rotating inner cylinder 3 of outer barrel 2；
Bottom in the inner cylinder 3 is provided with the first rabbling mechanism, and first rabbling mechanism is stirred including the first agitating shaft 4 and first
Mix blade 5, first agitating shaft 4 along the central shaft longitudinal direction of the inner cylinder 3 through inner cylinder 3 inside, described first stirs
Mix blade 5 and be connected with the first agitating shaft 4 and driven by the first agitating shaft 4 and rotated；Between the outer barrel 2 and inner cylinder 3
Interlayer in be additionally provided with the first motor, weigh 6 and first heating film；First motor drives the inner cylinder 3 to rotate,
And first agitating shaft 4 is driven to rotate；6 lower sections for being arranged at the inner cylinder 3 of weighing, with inner cylinder 3
Material is weighed；First heating film is covered on the outer wall of the inner cylinder 3, to be heated to the inner cylinder 3；
Filtrate port 7 is provided with the lower wall of the inner cylinder 3, the first filter screen is removably provided with the filtrate port 7.
Extract chamber 8, its upper part opening, it is described extraction chamber 8 opening on be provided with lid 9, by it is described extraction chamber 8 be open
Covering；The extraction chamber 8 passes through pipeline 15 and 7 fluid communication of filtrate port；The extraction chamber 8 includes outer chamber 10 and installation
And can the relatively described rotating inner chamber body 11 of outer chamber 10 in the outer chamber 10；Bottom in the inner chamber body 11 is provided with
Second rabbling mechanism, second rabbling mechanism include the second agitating shaft 12 and the second agitating vane 13, second agitating shaft
12 along the central shaft longitudinal direction of the inner chamber body 11 through inside inner chamber body 11,13 and second agitating shaft of the second agitating vane
12 connections are simultaneously rotated by the drive of the second agitating shaft 12；It is additionally provided with interlayer between the outer chamber 10 and inner chamber body 11
Second motor and the second heating film；Second motor drives the inner chamber body 11 to rotate, and drives second agitating shaft 12
Rotation；Second heating film is covered on the outer wall of the inner chamber body 11, to be heated to the inner chamber body 11；In described
Filtering mouth 14 is provided with the lower wall of cavity 11, the second filter screen is removably provided with the filtering mouth 14.
In such scheme, flower of JINHUAKUI is put into inner cylinder, and the temperature in inner cylinder is adjusted by the first heating film, the
One motor drives the inner cylinder rotation, and then carries out constant temperature drying, the flower of JINHUAKUI after drying to flower of JINHUAKUI in rotation
It is placed again into after crushing in inner cylinder, weighing below inner cylinder weighs the flower of JINHUAKUI of crushing, and user is according to title
Weight result can be segmented the enzymolysis liquid for adding suitable cellulase and distilled water mixing into inner cylinder, and then the first motor drives
First rabbling mechanism is stirred the enzymolysis liquid in inner cylinder and flower of JINHUAKUI crushed material, while the first rabbling mechanism is located all the time
In stirring, and the first heating film needs to adjust the temperature in inner cylinder according to reaction so that flower of JINHUAKUI is in lasting stirring
And digested under conditions of constant temperature, wherein, its cell membrane can be destroyed using cellulase extraction flower of JINHUAKUI polysaccharide, favorably
In polysaccharide dissolution；Afterwards the first heating film heat up, by temperature adjustment in inner cylinder to 92 DEG C or so, to flower of JINHUAKUI enzymolysis liquid into
Row inactivation, inner cylinder is rotated to 4000r/min afterwards, and the enzymolysis liquid after inactivation is centrifuged, and finally opens the reaction chamber
Filtrate port, the filtrate of the enzymolysis mixed liquor after the centrifugation automatically flows into extraction intracavitary by pipeline；To the inner chamber body of extraction chamber
Ethanol is added in interior filtrate, covers lid, stands 12h or so, you can alcohol precipitation is carried out to filtrate, in the second motor driving afterwards
Cavity is rotated to 4000r/min, and the filtrate after alcohol precipitation is centrifuged, and opens filtering mouth, you can flow out the liquid after centrifugation
Extract outside chamber, and 95% ethanol, absolute ethyl alcohol and acetone are added sequentially in sediment and is washed, coordinate second in the process
Rabbling mechanism is stirred sediment, so that it mixes substantially uniformity with each cleaning solution, afterwards by the second heating film by
Cavity is heated to 60 DEG C, you can realizes that the flower of JINHUAKUI polysaccharide that constant temperature obtains extraction is dried.
Pass through the structure setting of extractor so that flower of JINHUAKUI realizes automation mechanized operation when extracting polysaccharide, reduces
Artificial participation, reduces labour cost, and avoids and replace utensil repeatedly, and extract pollutes during causing or what is wasted asks
Topic so that extraction process is safely controllable, and more convenient, reduces extraction cost.
In one preferred solution, scale is respectively arranged with the inner wall of the inner cylinder 3 and inner chamber body 11.
In such scheme, by setting scale on the inner wall of inner cylinder and inner chamber body, the steaming to addition can be facilitated
The amount of the liquid such as distilled water, ethanol carries out control, so that extraction process is more convenient.
In one preferred solution, sealing ring is provided with the lower surface outer rim of the lid 9, so that the lid 9 covers
When in the opening of the extraction chamber 8, sealing space is formed in the extraction chamber 8.
In such scheme, because alcohol precipitation process needs to carry out under closed environment, thus by the lower surface of lid
Edge set sealing ring, can ensure lid closure when extract intracavitary airtight space, and then ensure alcohol precipitation process it is smooth
Carry out, and then ensure the recovery rate and extraction quality of flower of JINHUAKUI polysaccharide.
In one preferred solution, the lower section of the inner cylinder 3 is additionally provided with the intake chute 16 of annular, and the intake chute 16 is set
Center concave arc downwards is set to, so that the intake chute 16 forms the accommodating space of upper opening, the intake chute 16
Set in the lower section of the filtrate port 7, and around the inner cylinder 3, the lower section of the intake chute 16 offers fluid guidance port 17, institute
State fluid guidance port 17 and extraction 8 fluid communication of chamber.
In such scheme, by the lower section of inner cylinder set intake chute so that inner cylinder after rotation, no matter filtrate
Which direction mouth rushes at, and filtrate homomergic flow enters in intake chute, and the fluid guidance port of intake chute and extraction chamber fluid communication so that filtrate
Extraction intracavitary can be smoothly flowed into, carries out the extraction of follow-up polysaccharide.
Embodiment
Experimental data
Single factor design
The single factor test of the enzyme concentration of progress cellulase assisted extraction flower of JINHUAKUI polysaccharide, enzymolysis time and solid-liquid ratio successively
Experiment, every group of experiment carries out at least three groups and repeats to test, to reduce experimental error.
Influence of the enzyme amount to flower of JINHUAKUI polysaccharide extract rate：5 parts of dried powder of 2g are measured, keep primary condition not
Become：50 DEG C, extraction 1h, solid-liquid ratio 1:60g/mL, sets different 0.6%, 0.8%, 1.0%, 1.2%, 1.4% (enzymes of enzyme concentration
With Golden flower mass ratio) carry out experiment of single factor.
The results are shown in Figure 1, under 0.6%, 0.8%, 1.0%, 1.2%, 1.4% experiment condition of enzyme concentration, obtains gold
Tree mallow polysaccharide extract rate is respectively 2.81%, 2.94%, 3.52%, 3.05%, 2.67%.Generally flower of JINHUAKUI polysaccharide carries
Take and take the lead in reducing after increase.Cellulose enzyme amount is increased to before 1.00%, and flower of JINHUAKUI Polyose extraction takes the lead in increasing；Enzyme amount increases to
When 1.00%, flower of JINHUAKUI polysaccharide extract rate reaches peak 3.52%；With Polyose extraction under the conditions of enzyme concentration 0.8%, 1.2%
Rate has differed 0.58%, 0.47% respectively, and significant difference (P ＜ 0.05) is all presented.After enzyme amount is more than 1.0%, Polyose extraction
Rate is not further added by, and presentation is decreased obviously trend, reduces by 0.47%, 0.38% successively.Cellulose when illustrating to add 1.00% enzyme amount
Enzyme amount has reached saturation state.Therefore it is response surface experimental design cellulase dosage central horizontal to choose 1.00% enzyme amount.
Influence of the enzymolysis time to flower of JINHUAKUI polysaccharide extract rate：5 parts of dried powder of 2g are measured, primary condition is：
Enzyme amount 1.0%, temperature 50 C, solid-liquid ratio 1:60g/mL.Keep primary condition constant：Set different enzymolysis time 0.5h, 1.0h,
1.5h, 2.0h, 2.5h carry out experiment of single factor.
The results are shown in Figure 2, and in the case where extracting 0.5h, 1.0h, 1.5h, 2.0h, 2.5h experiment condition, it is more to obtain flower of JINHUAKUI
Sugared recovery rate is followed successively by 1.76%, 3.11%, 2.32%, 2.56%, 2.97%, and analysis recovery rate obtains corresponding standard error
Respectively 0.01%, 0.08%, 0.19%, 0.32%, 0.06%.Time 0.5h is carried from enzyme and extends to 1.0h, and flower of JINHUAKUI is more
Sugar extraction takes the lead in substantially increasing；During enzymolysis time 1h, flower of JINHUAKUI polysaccharide extract rate reaches peak 3.11%, with extraction time
Polysaccharide extract rate has differed 1.32%, 0.79% respectively under the conditions of 0.5h, 1.5h, and significant difference (P ＜ 0.05) is all presented.Afterwards
Polysaccharide extract rate has slow ascendant trend again after continuous 1.5h, it may be possible to which, because after the time increases, the activity of cellulase is lower, right
Polysaccharide extract rate influences smaller, approximation water extraction method extraction polysaccharide during subsequent extracted, and then when polysaccharide extract rate is mainly extracted
Between influence.Therefore choose 1h and put forward time centre level for response surface experimental design enzyme.
Influence of the solid-liquid ratio to flower of JINHUAKUI polysaccharide extract rate：5 parts of dried powder of 2g are measured, primary condition is：Enzyme
Amount 1.0%, temperature 50 C, extraction 1h.Keep primary condition constant：Different feed liquid is set than 1:50g/mL、1:60g/mL、1:
70g/mL、1:80g/mL、1:90g/mL carries out experiment of single factor.
The results are shown in Figure 3, in solid-liquid ratio 1:50g/mL、1:60g/mL、1:70g/mL、1:80g/mL、1:90g/mL is real
Under the conditions of testing, it is respectively 2.61%, 3.31%, 2.85%, 2.89%, 2.71% to obtain flower of JINHUAKUI polysaccharide extract rate, analysis
Recovery rate obtains corresponding standard error and is followed successively by 0.17%, 0.27%, 0.15%, 0.21%, 0.29%.With solid-liquid ratio
Increase, generally flower of JINHUAKUI Polyose extraction take the lead in reducing after increase.Solid-liquid ratio is from 1:50g/mL increases to 1:60g/mL, golden flower
The recovery rate of sunflower polysaccharide substantially increases；From 1:60g/mL to 1:70g/mL polysaccharide extract rates substantially reduce, from 1:70g/mL is arrived
1:90g/mL flower of JINHUAKUI polysaccharide extract rates do not have significant change, and trend is gentle.Solid-liquid ratio sets 1:During 60g/mL, Polyose extraction
Rate increases to peak 3.31%, with solid-liquid ratio 1:50g/mL、1:Flower of JINHUAKUI polysaccharide extract rate differs respectively under the conditions of 70g/mL
0.70%th, 0.46%, significant difference (P ＜ 0.05) is all presented.Solid-liquid ratio is more than 1:After 60g/mL, it may be possible to because solvent mistake
Cause cellulase concentration to reduce, polysaccharide extract rate declines more.Therefore 1 is chosen:60g/mL is response surface experimental design solid-liquid ratio
Central horizontal.
Response phase method experimental design
According to experiment of single factor as a result, selection enzyme amount, enzymolysis time, solid-liquid ratio are independent variable, carried with flower of JINHUAKUI polysaccharide
It is response to take rate, with Design Expert 8.0.6.1 softwares, is designed according to Box-Behnken principles, carries out three factors
Three horizontal response surface analysis experiments, analysis result are shown in Table 1, draw the optimal extraction process of flower of JINHUAKUI polysaccharide.
1 response surface experimental design factor of table and water-glass
According to table 1, Design Expert 8.0.6.1 softwares are run, the experiment of flower of JINHUAKUI Polyose extraction is carried out successively and sets
Meter, regression analysis and prediction optimization.A new design document is created first, and into response surface experimental design, input will be investigated
The number (3) of factor, title (enzyme amount, enzymolysis time, solid-liquid ratio), unit (%, h, g/mL) and factor it is horizontal up and down
It is worth (enzyme amount 3.05,2.94；Enzymolysis time 2.32,1.76；Solid-liquid ratio 2.85,2.61), rear to input response, that is, flower of JINHUAKUI more
Number (1), title (recovery rate) and the unit (%) of sugared recovery rate, draw flower of JINHUAKUI polysaccharide response surface designing scheme.Altogether into
17 groups of experiments of row, wherein sequence number 1,2,4,5,10 5 groups be center experiment, repeat five times to surmise experimental error.Remaining 12 groups real
It is factorial experiment to test.Experiment is carried out according to scheme and draws every group of polysaccharide extract rate, and in Input Software.Scheme and it the results are shown in Table 2.
The results of analysis of variance is shown in Table 3.
2 response surface experimental design of table and result
3 the results of analysis of variance of table
Note:* 0.05 significant differences of P ＜ are represented, * * represent that P 0.01 differences of ＜ are extremely notable.
Flower of JINHUAKUI polysaccharide extract rate data are analyzed, draw secondary multinomial regression equation：
Y=3.47-0.26A-0.44B-0.35C+0.092AB+0.053AC-0.16BC-0.66A2- 0.57B2-
0.89C2。
Y represents response flower of JINHUAKUI polysaccharide extract rate in formula, and A represents enzyme amount, and B represents enzymolysis time, and C represents liquid material
Than.By equation it can be seen that cellulose enzyme amount, enzymolysis time, three factors of liquid ratio and response flower of JINHUAKUI polysaccharide extract rate
It is not simple linear relationship.
A, C act on flower of JINHUAKUI polysaccharide extract rate notable, B, A in regression equation2、B2、C2To flower of JINHUAKUI Polyose extraction
Rate effect is extremely notable；AB, AC, BC be not notable to the effect of flower of JINHUAKUI polysaccharide extract rate, illustrates the interaction between these three factors
Effect is not notable.In three factors according to selected by being released the big I of F values in response surface experimental result and table 3, enzymolysis time (B) is right
The effect of flower of JINHUAKUI polysaccharide extract rate is most notable, and secondly liquid ratio (C) is more than enzyme to flower of JINHUAKUI polysaccharide extract rate influence degree
Measure the influence degree of (A).
The conspicuousness of the model is examined, the results of analysis of variance is shown in Table 3, and regression equation Analysis on confidence is shown in Table 4.
4 regression equation Analysis on confidence of table
The P values of model have 0.01,0.05 two boundary ,≤0.01 to represent that extremely notable ,≤0.05 represents notable, 0.05 tables of ＞
Show not notable.This model P=0.0018 as shown in Table 3<0.01, model is extremely notable.Lose plan item P values to be the bigger the better, as shown in Table 3
Lose and intend item P=0.2207>0.05, it is not significantly, with representing equation model relatively good.Total decision of the regression model as shown in Table 4
Coefficients R2=0.9389 ＞ 0.9, represents model close to actual conditions, R2It is better closer to 1；Adjust coefficient of determination R2 AdjEqually it is
Similar R2Measurement model a parameter, it can reduce influence of the multinomial number to related coefficient, this model R2 Adj=
0.8603, illustrate that the model experiment error is smaller.In summary, which can be applied to optimization cellulase assisted extraction golden flower
Sunflower polysaccharide extracting process.
Embodiment 1
In a kind of flower of JINHUAKUI extraction method of polysaccharides, mainly include the following steps：
Drying to constant weight being put into the reaction chamber being preheated at 60 DEG C in extractor after step 1, flower of JINHUAKUI are cleaned, and obtains
To after dry flower of JINHUAKUI, 60 mesh are crushed to, obtain Golden flower pollen；
Step 2, by obtained Golden flower pollen according to solid-liquid ratio 1:58.28g/mL weighs quantitative distilled water, Yi Jizhan
The cellulase of the Golden flower pollen gross mass 0.95%, temperature is mixed and heated to as 50 by the distilled water and cellulase
DEG C, add into the Golden flower pollen and stir evenly, it is 7 to adjust pH value, and lasting stirring enzymolysis 0.81h, obtains enzymolysis mixing
Liquid；
Step 3, after the enzymolysis mixed liquor is inactivated 10min at a temperature of 92 DEG C, centrifuge under the conditions of 4000r/min
10min, opens the filtrate port of the reaction chamber, and the filtrate of the enzymolysis mixed liquor after the centrifugation flows into extraction intracavitary；
Step 4, the volume according to filtrate in the extractor, the mass concentration that 4 times of volumes are added into the filtrate are
95% ethanol, sealing and standing 12h, obtains alcohol precipitation mixed liquor；
Step 5, by the alcohol precipitation mixed liquor centrifuge 15min under the conditions of 4000r/min, opens the filtering of the extractor
Mouthful, sediment is stayed in filtering；
Step 6, sequential purge and sink the sediment in ethanol that mass concentration is 95%, absolute ethyl alcohol and acetone
Form sediment, then constant temperature drying is to constant weight at 60 DEG C, up to the flower of JINHUAKUI polysaccharide.
Embodiment 2
In a kind of flower of JINHUAKUI extraction method of polysaccharides, mainly include the following steps：
Step 1, flower of JINHUAKUI clean after after 6 DEG C of Cord blood 4h, be put into and be preheated at 60 DEG C in the reaction of extractor
Intracavitary dries 1.5h, then reduces reaction chamber temperature and continues to dry 3h to 50 DEG C, finally reduces reaction chamber temperature to 35 DEG C of constant temperature
Drying to constant weight, after obtaining dry flower of JINHUAKUI, is crushed to 60 mesh, obtains Golden flower pollen；
Step 2, by obtained Golden flower pollen according to solid-liquid ratio 1:58.28g/mL weighs quantitative distilled water, Yi Jizhan
The cellulase of the Golden flower pollen gross mass 0.95%, 3 parts are divided into by the distilled water and cellulase respectively, and will
3 parts of distilled water are separately heated to temperature as 60 DEG C, 50 DEG C and 65 DEG C, and 3 parts of cellulases are separately added into 3 parts of distilled water
In to dissolve, obtain 3 portions of cellulase mixed liquors；3 portions of cellulase mixed liquors are then pressed into interval time 22min, are sequentially added
Enter into the Golden flower pollen and stir evenly, and it is 7 to adjust pH value all the time, lasting stirring, obtains enzymolysis mixed liquor；
Step 3, after the enzymolysis mixed liquor is inactivated 10min at a temperature of 92 DEG C, centrifuge under the conditions of 4000r/min
10min, opens the filtrate port of the reaction chamber, and the filtrate of the enzymolysis mixed liquor after the centrifugation flows into extraction intracavitary；
Step 4, the volume according to filtrate in the extractor, the mass concentration that 4 times of volumes are added into the filtrate are
95% ethanol, sealing and standing 12h, obtains alcohol precipitation mixed liquor；
Step 5, by the alcohol precipitation mixed liquor centrifuge 15min under the conditions of 4000r/min, opens the filtering of the extractor
Mouthful, sediment is stayed in filtering；
Step 6, sequential purge and sink the sediment in ethanol that mass concentration is 95%, absolute ethyl alcohol and acetone
Form sediment, then constant temperature drying is to constant weight at 60 DEG C, up to the flower of JINHUAKUI polysaccharide.
Embodiment 1 is that the optimal Extraction technique drawn using response phase method extracts flower of JINHUAKUI polysaccharide, and embodiment 2 is
The optimal Extraction technique drawn using response phase method and the drying of combination gradient and subsection enzymolysis extraction flower of JINHUAKUI polysaccharide.
Flower of JINHUAKUI polysaccharide is extracted using Examples 1 and 2 the method respectively, three groups is respectively done and repeats experiment to reduce experiment
Error, the flower of JINHUAKUI polysaccharide extract rate for drawing embodiment 1 is 3.58%, and model prediction flower of JINHUAKUI polysaccharide extract rate highest
For 3.6173%.Both relative errors are 1.04%, and less than critical value 5%, i.e., this model is to cellulase assisted extraction golden flower
The real result that the optimization of sunflower polysaccharide is drawn is credible, and the flower of JINHUAKUI polysaccharide extract rate of embodiment 2 is 4.02%, is significantly higher by
The polysaccharide extract rate of embodiment 1, it is seen that gradient dries the extraction efficiency for being conducive to flower of JINHUAKUI polysaccharide with the method for subsection enzymolysis
Raising.
Although embodiment of the present invention is disclosed as above, it is not restricted in specification and embodiment listed
With it can be applied to various suitable the field of the invention completely, can be easily for those skilled in the art
Realize other modification, therefore under the universal limited without departing substantially from claim and equivalency range, it is of the invention and unlimited
In specific details and shown here as the legend with description.
Claims (9)
1. a kind of flower of JINHUAKUI extraction method of polysaccharides, wherein, mainly include the following steps：
Step 1, flower of JINHUAKUI clean after after 5-10 DEG C of Cord blood 3-5h, be put into and be preheated at 55-65 DEG C in extractor
Drying 1-2h in reaction chamber, then reduces reaction chamber temperature and continues to dry 2-4h to 45-55 DEG C, finally reduce reaction chamber temperature extremely
30-45 DEG C of constant temperature drying after obtaining dry flower of JINHUAKUI, is crushed to 50-70 mesh, obtains Golden flower pollen to constant weight；
Step 2, by obtained Golden flower pollen according to solid-liquid ratio 1:50-60g/mL weighs quantitative distilled water, and accounts for described
The cellulase of Golden flower pollen gross mass 0.8-1.2%, 3 parts are divided into by the distilled water and cellulase respectively, and by 3
Part distilled water is separately heated to temperature as 30-45 DEG C, 45-50 DEG C and 60-65 DEG C, and 3 parts of cellulases are separately added into 3
To dissolving in part distilled water, 3 portions of cellulase mixed liquors are obtained；3 portions of cellulase mixed liquors are then pressed into interval time 20-
30min, is added sequentially into the Golden flower pollen and stirs evenly, and adjusts pH value all the time between 6.5-7.5, persistently stirs
Mix, obtain enzymolysis mixed liquor；
Step 3, after the enzymolysis mixed liquor is inactivated 8-12min at a temperature of 85-100 DEG C, in 3800-4200r/min bars
8-12min is centrifuged under part, opens the filtrate port of the reaction chamber, is carried described in the filtrate inflow of the enzymolysis mixed liquor after the centrifugation
Take the extraction intracavitary of device；
Step 4, the volume according to filtrate in the extractor, the mass concentration that 3-5 times of volume is added into the filtrate is 92-
96% ethanol, sealing and standing 10-15h, obtains alcohol precipitation mixed liquor；
Step 5, by the alcohol precipitation mixed liquor centrifuge 10-20min under the conditions of 3800-4200r/min, opens the extractor
Mouth is filtered, sediment is stayed in filtering；
Step 6, sequential purge and sink the sediment in mass concentration is the ethanol of 92-96%, absolute ethyl alcohol and acetone
Form sediment, then constant temperature drying is to constant weight at 55-65 DEG C, up to the flower of JINHUAKUI polysaccharide.
2. flower of JINHUAKUI extraction method of polysaccharides as claimed in claim 1, wherein, mainly include the following steps：
Step 1, flower of JINHUAKUI clean after after 6 DEG C of Cord blood 4h, be put into and be preheated at 60 DEG C in the reaction chamber of extractor
1.5h is dried, reaction chamber temperature is then reduced and continues to dry 3h to 50 DEG C, finally reduce reaction chamber temperature to 35 DEG C of constant temperature dryings
To constant weight, after obtaining dry flower of JINHUAKUI, 60 mesh are crushed to, obtain Golden flower pollen；
Step 2, by obtained Golden flower pollen according to solid-liquid ratio 1:58.28g/mL weighs quantitative distilled water, and accounts for described
The cellulase of Golden flower pollen gross mass 0.95%, 3 parts are divided into by the distilled water and cellulase respectively, and by 3 parts
Distilled water is separately heated to temperature as 60 DEG C, 50 DEG C and 65 DEG C, and 3 parts of cellulases are separately added into 3 parts of distilled water
To dissolving, 3 portions of cellulase mixed liquors are obtained；3 portions of cellulase mixed liquors are then pressed into interval time 22min, are added sequentially
Stirred evenly into the Golden flower pollen, and it is 7 to adjust pH value all the time, lasting stirring, obtains enzymolysis mixed liquor；
Step 3, after the enzymolysis mixed liquor is inactivated 10min at a temperature of 92 DEG C, centrifuge under the conditions of 4000r/min
10min, opens the filtrate port of the reaction chamber, and the filtrate of the enzymolysis mixed liquor after the centrifugation flows into extraction intracavitary；
Step 4, the volume according to filtrate in the extractor, the mass concentration that 4 times of volumes are added into the filtrate are 95%
Ethanol, sealing and standing 12h, obtains alcohol precipitation mixed liquor；
Step 5, by the alcohol precipitation mixed liquor centrifuge 15min under the conditions of 4000r/min, opens the filtering mouth of the extractor,
Sediment is stayed in filtering；
Step 6, sequential purge and precipitate the sediment in ethanol that mass concentration is 95%, absolute ethyl alcohol and acetone,
Then constant temperature drying is to constant weight at 60 DEG C, up to the flower of JINHUAKUI polysaccharide.
3. flower of JINHUAKUI extraction method of polysaccharides as claimed in claim 2, wherein, after dry flower of JINHUAKUI is crushed in step 1
Cross 60 mesh sieves.
4. flower of JINHUAKUI extraction method of polysaccharides as claimed in claim 2, wherein, the mesh of the filter screen of filtrate port in step 3
For 65-80 mesh.
5. flower of JINHUAKUI extraction method of polysaccharides as claimed in claim 2, wherein, the filtering of the filtering mouth of extractor in step 5
The mesh of net is 70-95 mesh.
6. a kind of extractor applied to flower of JINHUAKUI extraction method of polysaccharides as claimed in claim 1, wherein, including：
Reaction chamber, its upper part opening, the reaction chamber include outer barrel and in the outer barrel and can relatively it is described outside
The rotating inner cylinder of cylinder；Bottom in the inner cylinder is provided with the first rabbling mechanism, and first rabbling mechanism includes the
One agitating shaft and the first agitating vane, first agitating shaft is along the central shaft longitudinal direction of the inner cylinder through in inner cylinder
Portion, first agitating vane are rotated with the first stirring axis connection and by the drive of the first agitating shaft；The outer barrel and interior
The first motor is additionally provided with interlayer between cylinder, is weighed and the first heating film；First motor drives the inner cylinder
Body rotates, and drives first agitating shaft to rotate；Described weigh is arranged at the lower section of the inner cylinder, with inner cylinder
Material weigh；First heating film is covered on the outer wall of the inner cylinder, to be heated to the inner cylinder；
Filtrate port is provided with the lower wall of the inner cylinder, the first filter screen is removably provided with the filtrate port；
Extract chamber, its upper part opening, it is described extraction chamber opening on be provided with lid, by it is described extraction chamber opening cover；
The extraction chamber passes through pipeline and the filtrate port fluid communication；The extraction chamber includes outer chamber and installed in the outer chamber
It is interior and can the relatively described rotating inner chamber body of outer chamber；Bottom in the inner chamber body is provided with the second rabbling mechanism, and described
Two rabbling mechanisms include the second agitating shaft and the second agitating vane, central shaft longitudinal direction of second agitating shaft along the inner chamber body
Through inside inner chamber body, second agitating vane is rotated with the second stirring axis connection and by the drive of the second agitating shaft；
The second motor and the second heating film are additionally provided with interlayer between the outer chamber and inner chamber body；Second motor drives institute
Inner chamber body rotation is stated, and drives second agitating shaft to rotate；Second heating film is covered on the outer wall of the inner chamber body,
To be heated to the inner chamber body；It is provided with filtering mouth in the lower wall of the inner chamber body, it is dismountable on the filtering mouth
It is provided with the second filter screen.
7. extractor as claimed in claim 6, wherein, it is respectively arranged with scale on the inner wall of the inner cylinder and inner chamber body.
8. extractor as claimed in claim 6, wherein, sealing ring is provided with the lower surface outer rim of the lid, so that institute
State lid be covered in it is described extraction chamber opening on when, it is described extraction intracavitary formed sealing space.
9. extractor as claimed in claim 6, wherein, the intake chute of annular is additionally provided with below the inner cylinder, it is described
Intake chute downward concave arc centered on setting, so that the intake chute forms the accommodating space of upper opening, the drain
Groove is located at the lower section of the filtrate port, and is set around the inner cylinder, and fluid guidance port is offered below the intake chute, described
Fluid guidance port and the extraction chamber fluid communication.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110256592A (en) * | 2019-06-25 | 2019-09-20 | 金坤石家庄众创空间有限公司 | A kind of extraction element and method for Golden flower polysaccharide |
| CN110537686A (en) * | 2019-10-10 | 2019-12-06 | 陕西科技大学 | Selenium-rich black garlic and abelmoschus manihot chewable tablets and preparation method thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110256592A (en) * | 2019-06-25 | 2019-09-20 | 金坤石家庄众创空间有限公司 | A kind of extraction element and method for Golden flower polysaccharide |
| CN110537686A (en) * | 2019-10-10 | 2019-12-06 | 陕西科技大学 | Selenium-rich black garlic and abelmoschus manihot chewable tablets and preparation method thereof |
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Application publication date: 20180511 |